Full sequencing of the BRCA1/2 genes, plus deletion/duplication analysis. May be used to assess the risk of carrying a BRCA1/2 mutation when there is no known familial mutation.
According to the National Comprehensive Cancer Network®1, testing is indicated if one of the features mentioned below is present in the family: Early-age-onset (age <50 years) breast cancer, including both invasive and ductal carcinoma in situ (DCIS) breast cancers; two breast primaries or breast and ovarian/fallopian tube/primary peritoneal cancer in a single individual or two or more breast primaries or breast and ovarian/fallopian tube/primary peritoneal cancers in close (first-, second-, and third-degree) relatives(s) from the same side of the family; populations at risk (eg, Ashkenazi Jewish); member of a family with a known BRCA1 or BRCA2 mutation; any male breast cancer; ovarian/fallopian tube/primary peritoneal cancer at any age.
Next generation sequencing and multiplex ligation-dependent probe amplification (MLPA) platform
Lavender-top (EDTA) tube or yellow-top (ACD) tube
A BRCAssure clinical questionnaire should be submitted with specimens. Contact CMBP genetics services at 800-345-4363 to coordinate testing.
Causes for Rejection
Frozen whole blood, serum or marrow; leaking tube; clotted blood or marrow; grossly hemolyzed specimen; incorrect anticoagulant
Maintain specimen at room temperature
The entire gene coding region of BRCA1/BRCA2 genes, as well as all flanking noncoding regions, is analyzed by next generation sequencing. The test uses Illumina's TruSeq Custom Amplicon approach for target enrichment and MiSeq instrument for sequencing. It is a custom designed pool comprised of 150 oligonucleotide probe pairs for the binding and amplification of products ranging from 220bp to 280bp. Sequencing does not detect inversions, or other rearrangements.
Comprehensive deletion/duplication testing for BRCA1/BRCA2 is performed by using the multiplex ligation-dependent probe amplification (MLPA) platform. The MLPA will detect full BRCA1/BRCA2 gene deletions and multiple- and single-exon deletions and duplications. Balanced translocations are not detected by the method.